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本实验所用的中国红豆杉细胞悬浮培养体系中,云南紫杉烷c(Tc)是主要的次生代谢产物,该化合物有类神经生长因子活性,提高其产量是进一步规模化生产的前提。本研究考察了原位吸附和茉莉酸甲酯(MJA)联合调控提高Tc产量的可能性。在培养的第7天加入浓度为100μmol/L的MJA虽然会使细胞的生物量下降10%～30%,但是单位细胞内Tc含量和Tc产量均有显著提高,分别是对照的3.6和3.3倍。吸附剂XAD-7在不同时间加入对Tc的合成影响显著。在培养的第7天同时加入100μmol/L的MJA和100g/L的XAD-7会使细胞生物量增加,Tc产量显著提高。培养到第21天,Tc产量达477.4mg/L,为对照的6.3倍,为只加MJA的1.9倍,其中94%的Tc被树脂吸附。实验结果表明,在MJA诱导高表达的过程中,吸附剂XAD-7的加入使细胞内代谢产物外泌,浓度降低,减轻产物反馈抑制现象,从而大幅度提高代谢物产量,有较好的生产前景。
Taxus c (Tc) in Yunnan Taxus cell suspension culture system used in this experiment is the main secondary metabolites, the compound has the activity of nerve growth factor to increase its production is the premise of further large-scale production. This study investigated the possibility of in situ adsorption and methyl jasmonate (MJA) regulation to increase Tc production. Adding MjA at the concentration of 100μmol / L on the 7th day of culture could reduce the biomass of the cells by 10% -30%, but the Tc content and Tc production per unit cell increased significantly, which were 3.6 and 3.3 times of the control respectively . Addition of adsorbent XAD-7 at different times significantly affected the synthesis of Tc. The addition of 100μmol / L MJA and 100g / L XAD-7 on the 7th day of culture resulted in an increase of cell biomass and a significant increase of Tc production. Tc production reached 477.4mg / L on the 21st day, which is 6.3 times that of the control, which is 1.9 times of that of the MJA alone, and 94% of the Tc is adsorbed by the resin. The results showed that in the process of MJA induced high expression, the addition of adsorbent XAD-7 reduced the exogenous secretion and concentration of metabolites in the cells and reduced the feedback inhibition of products, thus greatly improving the yield of metabolites and better production prospect.